Lecture: Enzyme03

1. Enzymes Pt 3: Regulation of enzyme activity Relicardo M. Coloso, Ph. D. College of Medicine Central Philippine University

2. Regulate – to control or direct according to a rule, principle or law Enzyme regulation- is the control of the rate of a reaction catalyzed by an enzyme by some effector (e.g., inhibitors or activators) or by alteration of some condition (e.g., pH or ionic strength).

3. Five ways by which enzyme activity in the cell can be changed: 1. Enzyme production – synthesis or degradation 2. Compartmentation – different metabolic pathways occur in different cell compartments 3. Activation and inhibition – by activators or inhibitors, for example feed back inhibition by one of products of the reaction 4. Post-translational modification – for example by phosphorylation, methylation, glycosylation 5. Localization to a different environment – from a reducing (cytoplasm) to an oxidizing (periplasm) environment, of high pH to a low pH, or low salinity to high salinity, high to low energy charge

6. Mechanisms of Regulation

10. Cooperativity shown by an allosteric enzyme

13. Allosteric regulation of aspartate transcarbamoylase (ATCase), the initial enzyme in synthesis of pyrimidines This enzyme comprises a pair of trimeric catalytic subunits (orange) connected by three pairs of dimeric regulatory subunits (green). Binding of cytidine triphosphate (CTP; the blue dot ) to the regulatory subunits causes a conformational transition from the active R state to the inactive T state. The more open conformation of the R state permits substrate binding. Thus an increase in the concentration of CTP, an end product in the pyrimidine pathway, shuts off ATCase, an example of feedback inhibition. Copyright © 2000, W. H. Freeman and Company Allosteric transition between active and inactive states

15. Cyclic phosphorylation and dephosphorylation is a common cellular mechanism for regulating protein activity In this example, the target protein R (orange) is inactive when phosphorylated and active when dephosphorylated; the opposite pattern occurs in some proteins . Copyright © 2000, W. H. Freeman and Company Phosphorylation and dephosphorylation

16. A linear representation of the conversion of chymotrypsinogen into chymotrypsin by the excision of two dipeptides These reactions yield three separate chains (A, B, and C), which are covalently linked by disulfide bonds (yellow) in the active enzyme . In the folded, native conformation of chymotrypsin, histidine 57, aspartate 102, and serine 195 are located in the active site. Copyright © 2000, W. H. Freeman and Company Proteolytic activation

17. Thank you!